摘要目的 探讨腺病毒36型(Ad36)在诱导3T3-L1细胞棕色化过程中的作用.方法 对鸡尾酒法诱导组(对照组)和鸡尾酒加Ad36诱导组(实验组)第0、2、4、6、8天3T3-L1细胞进行BODIPY染色,观察细胞成脂情况;实时荧光定量PCR和免疫印迹法检测2组解耦联蛋白1(Ucp1)、ATP 合成酶 O亚基(Atp5o)、细胞色素 c 氧化酶亚基 5B ( Cox5b)、周脂素( Perilipin)的 mRNA 与蛋白表达水平.结果 BODIPY染色结果 显示,随着诱导天数的增加,对照组脂肪细胞内脂滴呈现出由小到大的动态变化,而实验组细胞内脂滴先增大,在第4天加入Ad36后脂滴变小.与对照组相比,实验组Ucp1、Atp5o、Cox5b mRNA和蛋白表达水平显著升高,Perilipin mRNA和蛋白表达水平则明显下降(P＜0.05).结论 在3T3-L1细胞分化过程中,Ad36通过正向调控Ucp1、Atp5o、Cox5b基因表达促进3T3-L1细胞棕色化.

abstractsObjective To investigate the role of adenovirus type 36 ( Ad36) in the browning of 3T3-L1 cells. Methods BODIPY staining was performed on the 0, 2nd, 4th, 6th and 8th days of the cocktail induction (control) group and the cocktail plus Ad36 induction ( experimental) group to observe the adipogenesis of 3T3-L1 cells. The mRNA and protein expressions of uncoupling protein-1 ( Ucp1 ), ATP synthase, H+ transporting, mitochondrial F1 complex, O subunit ( Atp5o), cytochrome c oxidase subunit 5B ( Cox5b), and perilipin were detected by real-time PCR and Western-blot. Results The results of BODIPY staining showed that the lipid droplets in the control group gradually became larger with the differentiation of the cells, while the lipid droplets in the experimental group firstly became larger, and then appeared smaller after Ad36 was added on the fourth day. Compared with the control group, the mRNA and protein expression levels of Ucp1, Atp5o, and Cox5b in the experimental group were significantly increased while the expression level of perilipin was significantly decreased ( all P＜0. 05 ). Conclusion During the differentiation of 3T3-L1 cells, Ad36 promotes its browning via increasing the expressions of Ucp1, Atp5o, and Cox5b.